An improved synthesis of a fluorophosphonate–polyethylene glycol–biotin probe and its use against competitive substrates

• Hao Xu,
• Hairat Sabit,
• Gordon L. Amidon and
• H. D. Hollis Showalter

Beilstein J. Org. Chem. 2013, 9, 89–96, doi:10.3762/bjoc.9.12

• these competition studies demonstrate a new application of FP-based probes seldom explored before. Keywords: biotin; fluorophosphonate; high turnover rate; reversible substrate; Introduction One of the goals of chemical biology is to develop small–molecule- and biomolecule-based probes to interrogate

• by the Cravatt group [21]. Kinetic study of FP probe labelling activity An incubation-time control study is the key toward successfully monitoring the competition between FP probe 1 and a reversible substrate with a high turnover rate. Hence, over a long incubation time, the substrate will be

• human tissues, is shown to be active in the hydrolysis of both enalapril and oseltamivir [23][24]. Each is hydrolyzed at a high turnover rate. In a control run, pCES (± preheated) was treated with 4 μM FP–PEG–biotin 1 (Figure 4A, lanes 1 and 2). Then in subsequent runs, non-preheated samples of enzyme